JP2005102501A - Cell culture apparatus and method for measuring dose responsiveness - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell culture apparatus designed to accurately measure the dose responsiveness of cells with different quantities of current even when the properties and number of the cells divided and injected in periods of experiments are different. <P>SOLUTION: The cell culture apparatus is composed so as to arrange permeable membranous membranes 23, 25 and 27 having different areas in series relatively to a current system as a means for producing current densities in order to apply at least the two different current densities to the cells in a culture medium in a vessel 16. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a cell culture apparatus and a dose response measuring method for culturing cells in a medium.
[0002]
[Prior art]
In general, in order to measure the dose response of a cell due to a direct current or an induced current, a cell having a different capacity is applied to the cell, or a different electric field is applied to cause a different capacity of the induced current to flow. Measure the difference in response to. Responsiveness includes changes in the number of cells, changes in cell genes, changes in cell chromosomes, changes in cell functions, changes in cell components, and the like. In general, culture dishes, culture cages, and culture membranes are used as cell retention systems for investigating responsiveness, but culture dishes of the same area, culture cages of the same area, and culture membranes of the same area are used. Yes. The cells are usually cultured in a culture dish or the like, and the cells taken out from the cells are diluted, and then a predetermined amount is separated and put into a cell holding system. This means that each time a different dose experiment is performed, cells are put into the retention system. Then, different currents are passed at different times, and the amount of change is measured.
[0003]
[Problems to be solved by the invention]
In the above method, the experiment period is divided for each different dose. Unless it is a cell at the same time, the nature of the cell and the number of cells to be compared for the dose cannot be confirmed. The reason is that, as time passes, there is a possibility that the cultured cells will grow or change their properties under ambient conditions. Therefore, the responsiveness of the cell due to the dose by different cells is measured, and inaccuracy is unavoidable. Therefore, conventionally, in order to eliminate the above drawbacks, many laboratory instruments are used to flow different amounts of electricity, and the same cells held in many holding systems at the same time are incorporated into the laboratory instruments. It was requested to start the experiment.
[0004]
The present invention has been made in view of the above-described drawbacks, and the object of the present invention is to provide a dose response of cells with different amounts of current even if the nature of the cells to be injected is divided and the number of cells is different. An object of the present invention is to provide a cell culture device and a dose responsiveness measuring method capable of accurately measuring sex.
[0005]
[Means for Solving the Problems]
A cell culture device according to claim 1, wherein a container for culturing cells in a medium by applying a predetermined voltage between a pair of electrodes installed at opposite ends and causing a current to flow between the electrodes; Current density generating means for providing at least two different current densities to cells in the medium in the container, and the current density generating means is arranged in series with the flowing current system.
[0006]
A cell culture device according to claim 2 is a container for culturing cells in a medium by applying a predetermined voltage between a pair of electrodes installed at opposite ends and causing a current to flow between the electrodes, Current density generating means for providing at least two different current densities to the cells in the medium in the container, the current density generating means being arranged in series with the flowing current system, the current density An electrically permeable barrier member disposed between the generating means and the electrode.
[0007]
A cell culture device according to claim 3 is the cell culture device according to claim 1 or 2, wherein the current density generating means is formed of a permeable main brain membrane having an ultra-micro structure in which cells cannot enter and exit, The brain film is provided on a holding body disposed at a predetermined interval.
[0008]
The dose responsiveness measuring method according to claim 4 includes a step of applying a predetermined voltage between a pair of electrodes installed at opposite ends and flowing a current between the electrodes, and at least the cells in the medium at the same time. Providing two different current densities.
[0009]
Embodiment
Hereinafter, one example of a cell culture device concerning the present invention is explained in full detail with reference to an accompanying drawing. FIG. 1 is an outline of a cell culture apparatus. Reference numeral 10 denotes a cell culture device. The cell culture device includes a power source 12 and a current control device 14 for controlling a current from the power source. The cell culture device is connected in series to an electrode of the cell culture device, and a predetermined amount of current flows in a closed circuit.
[0010]
The cell culture device 10 includes a container 16 having a rectangular parallelepiped inner hollow predetermined volume and capable of holding a fluid. The container is provided with electrodes (18, 18 ') on opposite ends (inner walls) thereof. And the culture medium for measuring the responsiveness of a cell is put in a container. In the container 16, a plurality of pairs, for example, three pairs of holding bodies (22, 24, 26) are arranged with a predetermined interval between both electrodes. The plurality of holding bodies are respectively arranged in series toward the system through which current flows.
[0011]
The holding body (22, 24, 26) is provided with a permeable main brain membrane (23, 25, 27) having an ultra-micro structure in which current permeability is good and cells cannot enter and exit. The permeable main brain membrane has a mesh diameter of about 0.3 mm and a thickness of about several to 10 microns.
[0012]
Then, an electrically permeable barrier (28, 28 ') such as agarose is disposed between the container and the holding member. Agarose may be applied to the surface of each electrode. By providing agarose, the influence of the chemical reaction in the vicinity of the electrode surface on the cells can be reduced.
[0013]
The holding body (22, 24, 26) includes a permeable main brain film only in a portion where current flows. That is, a substantially circular permeable main brain membrane (23, 25, 27) is provided only at a predetermined position where current flows between both electrodes. The circular area of the permeable main brain film is set to the diameters d1, d2, and d3 so as to be different for the three pairs of holding bodies. For example, if the circular area ratio of the permeable main brain film is 1: 2: 3, the current density is 1/3, 1/2, and 1.
[0014]
A medium containing cells is placed between the holding bodies arranged at a predetermined interval. Thereafter, a predetermined voltage is applied between both electrodes to pass a current. The current density in the medium can be obtained by dividing the value of the flowed current by the area of the permeable main brain membrane. Then, a predetermined voltage is applied between a pair of electrodes placed at opposite ends to cause a current to flow between the electrodes, so that at least two different current densities are given to cells in the medium at the same time, and dose response Measure.
[0015]
In the above embodiment, the pair of electrodes are arranged in the left-right direction and the direction of current flow is set to the horizontal direction. However, the pair of electrodes may be arranged in the up-down direction and the direction of current flow may be set to the vertical direction. By flowing the current in the vertical direction, it becomes possible to flow the current more stably and uniformly to the cells.
[0016]
【The invention's effect】
According to the cell culture device and the dose responsiveness measuring method of the present invention, the dose responsiveness of cells with different amounts of current can be accurately determined even when the experiment time is divided and the properties and the number of cells to be injected are different. Can be measured.
[Brief description of the drawings]
FIG. 1 is a schematic view showing a cell culture apparatus according to the present invention.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 10 Cell culture apparatus 12 Power supply 14 Current control apparatus 16 Container 18,18 'Electrode 22,24,26 Holding body 23,25,27 Permeable main brain film | membrane 28,28' Electropermeable barrier member

Claims (4)

A container for culturing cells in a medium by applying a predetermined voltage between a pair of electrodes placed at opposite ends and passing a current between the electrodes;
Current density generating means for providing at least two different current densities to cells in the medium in the container,
The current density generating means is arranged in series with the flowing current system;
A cell culture device. A container for culturing cells in a medium by applying a predetermined voltage between a pair of electrodes placed at opposite ends and passing a current between the electrodes;
Current density generating means for providing at least two different current densities to cells in the medium in the container,
The current density generating means is arranged in series with the flowing current system;
A cell culture device comprising: an electropermeable barrier member disposed between the current density generating means and the electrode.   The current density generating means is formed of a permeable main brain membrane having an ultra-micro structure in which cells cannot enter and exit. The cell culture device according to claim 1 or 2, wherein   A step of applying a predetermined voltage between a pair of electrodes disposed at opposite ends to cause a current to flow between the electrodes, and a step of applying at least two different current densities to cells in the medium at the same time. A dose responsiveness measuring method characterized by the above.

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